DNA损伤应答(DNAdamageresponse,DDR)机制包括检测DNA损伤,阻滞细胞周期和启动DNA修复。共济失调毛细血管扩张和Rad3相关激酶(ataxia telangiectasia and Rad3-related,ATR)是DDR核心的关键激酶,负责感知复制应激(replica-tion stress,...DNA损伤应答(DNAdamageresponse,DDR)机制包括检测DNA损伤,阻滞细胞周期和启动DNA修复。共济失调毛细血管扩张和Rad3相关激酶(ataxia telangiectasia and Rad3-related,ATR)是DDR核心的关键激酶,负责感知复制应激(replica-tion stress,RS)并将其信号传导至S和G2/M检查点以启动DNA修复。在肿瘤细胞中G1检查点缺失和癌基因的激活,导致癌症细胞更多进入RS增加的S期。因此,肿瘤细胞更加依赖S和G2/M检查点,使其成为一个有吸引力的靶点。ATR抑制剂是目前抗肿瘤药物开发的热点,部分ATR抑制剂目前已经进入临床试验阶段。本综述旨在总结支持ATR抑制剂作为单药以及与化疗、放疗和新型靶向药物(如PARP抑制剂)联合使用的临床试验数据,并讨论目前ATR抑制剂开发和生物标志物探索中面临的挑战。展开更多
Objective:Anlotinib hydrochloride is a multitarget tyrosine kinase inhibitor that targets vascular endothelial growth factor receptor,fibroblast growth factor receptor,platelet-derived growth factor receptor,c-Kit,and...Objective:Anlotinib hydrochloride is a multitarget tyrosine kinase inhibitor that targets vascular endothelial growth factor receptor,fibroblast growth factor receptor,platelet-derived growth factor receptor,c-Kit,and c-MET;therefore,it exhibits both antitumor and anti-angiogenetic activities.A phase III trial has shown that anlotinib improved progression-free survival(PFS)and overall survival(OS)in patients with advanced non-small cell lung cancer(NSCLC),who presented with progressive disease or intolerance after standard chemotherapy.This study aimed to analyze the characteristics of patients receiving anlotinib treatment to determine the dominant populations who are fit for the treatment.Methods:Data were collected from March 2015 to January 2017 from a randomized,double-blind,placebo-controlled,multicenter,phase III trial of anlotinib(ALTER0303).A total of 437 patients were enrolled and randomly allocated(2:1)to the anlotinib and placebo groups.Kaplan–Meier analysis and log-rank test were performed to compare PFS and OS.Cox proportional hazards model was adopted for multivariate prognostic analysis.Results:Multivariate analysis indicated that high post-therapeutic peripheral blood granulocyte/lymphocyte ratio and elevated alkaline phosphatase levels were independent risk factors for PFS.Meanwhile,elevated thyroid-stimulating hormone,blood glucose,and triglyceride levels;hypertension;and hand–foot syndrome were independent protective factors of PFS.High posttherapeutic peripheral blood granulocyte/lymphocyte ratio,an Eastern Cooperative Oncology Group(ECOG)score≥2,and the sum of the maximal target lesion length at baseline were independent risk factors of OS,and hypertriglyceridemia was an independent protective factor of OS.Conclusions:This study preliminarily explored the possible factors that affected PFS and OS after anlotinib treatment in patients with advanced refractory NSCLC,and the baseline characteristics of the therapeutically dominant populations were then identified.展开更多
Objective:Lymphatic metastasis is one of the leading causes of malignancy dispersion in various types of cancer.However,few anti-lymphangiogenic drugs have been approved for clinical use to date.Therefore,new therapie...Objective:Lymphatic metastasis is one of the leading causes of malignancy dispersion in various types of cancer.However,few anti-lymphangiogenic drugs have been approved for clinical use to date.Therefore,new therapies to block lymphangiogenesis are urgently required.Methods:Immunohistochemistry,immunofluorescence,Western blot,migration assays,and lymphangiogenesis and lymphatic metastasis assays were used.Results:Anlotinib,a receptor tyrosine kinase inhibitor,suppressed the rate of new metastatic lesions(31.82%in the placebo arm and 18.18%in the anlotinib arm)in patients with advanced lung adenocarcinoma who were enrolled in our ALTER-0303 study.D2-40+-lymphatic vessel density was strongly correlated with disease stage,metastasis,and poor prognosis in 144 Chinese patients with lung adenocarcinoma.In mice bearing A549EGFP tumors,tumor lymphatic vessel density,tumor cell migration to lymph nodes,and the number of distant metastatic lesions were lower in the anlotinib group than in the controls.Anlotinib inhibited the growth and migration of human lymphatic endothelial cells(hLECs)and lymphangiogenesisin vitro andin vivo.Treatment of hLECs with anlotinib downregulated phosphorylated vascular endothelial growth factor receptor 3(VEGFR-3).Conclusions:Anlotinib inhibits lymphangiogenesis and lymphatic metastasis,probably through inactivating VEGFR-3 phosphorylation.The results indicate that anlotinib may be beneficial for treatment in avoiding lymphangiogenesis and distant lymphatic metastasis in lung adenocarcinoma.(Trial registration:ALTER0303;NCT02388919;March 17,2015.)展开更多
Objective:Bevacizumab is a recombinant humanized monoclonal antibody that blocks vascular endothelial growth factor(VEGF)with clear clinical benefits.However,overall survival of some cancer types remains low owing to ...Objective:Bevacizumab is a recombinant humanized monoclonal antibody that blocks vascular endothelial growth factor(VEGF)with clear clinical benefits.However,overall survival of some cancer types remains low owing to resistance to bevacizumab therapy.While resistance is commonly ascribed to tumor cell invasion induced by hypoxia-inducible factor(HIF),less attention has been paid to the potential involvement of endothelial cells(ECs)in vasculature activated by anti-angiogenic drugs.Methods:Human umbilical vein ECs(HUVECs),bEnd.3 cells,and mouse retinal microvascular ECs(MRMECs)were treated with bevacizumab under conditions of hypoxia and effects on biological behaviors,such as migration and tube formation,examined.Regulatory effects on TGFpi and CD 105(endoglin)were established via determination o f protein and mRNA levels.We further investigated whether the effects of bevacizumab could be reversed using the receptor tyrosine kinase inhibitor anlotinib.Results:Bevacizumab upregulated TGFpi as well as CD 105,a component o f the TGFP receptor complex and an angiogenesis promoter.Elevated CD 105 induced activation of Sm adl/5,the inflammatory pathway and endothelial-mesenchymal transition.The migration ability of HUVECs was enhanced by bevacizumab under hypoxia.Upregulation o f CD 105 was abrogated by anlotinib,which targets multiple receptor tyrosine kinases including VEGFR2/3,FGFR1-4,PD G FRα/β,C-Kit,and RET.Conclusions:Bevacizumab promotes migration and tube formation of HUVECs via activation of the TGFβi pathway and upregulation of CD105 expression.Anlotinib reverses the effects of bevacizumab by inhibiting the above signals.展开更多
Determination of Cas9 cutting efficiency to the target sites is important for genome editing.However,this determination can only be made via an in vitro method,as the purification of Cas protein and synthesis of gRNA ...Determination of Cas9 cutting efficiency to the target sites is important for genome editing.However,this determination can only be made via an in vitro method,as the purification of Cas protein and synthesis of gRNA are necessary.Here,we developed an in vivo method,called transient CRISPR/Cas editing in plants(TCEP)to determine Cas9 cutting efficiency.The CRISPR/Cas vector for plant transformation mediated by Agrobacterium tumefaciens was constructed as normal.Using the transient transformation method we built,the Cas9 protein and gRNA were transiently expressed and formed a complex to cut its target sites,resulting in dynamic DNA breakage.The broken DNA was quantified using qPCR to measure the efficiencies of Cas9 cutting.We studied the Cas9 cutting efficiencies to different target sites in Betula platyphylla and Populus davidiana×P.bolleana plants using TCEP and an in vitro method.The results of TCEP were consistent with those of the in vitro method,suggesting that the TCEP method is reliable in determining cutting efficiency.Additionally,using the TCEP method,we showed that both heat and sonication treatment significantly improved CRISPR/Cas efficiency.Therefore,the TCEP method has broad application value and can not only be used to analyze the CRISPR/Cas efficiency but also to determine the factors involved in Cas9 cutting.展开更多
文摘DNA损伤应答(DNAdamageresponse,DDR)机制包括检测DNA损伤,阻滞细胞周期和启动DNA修复。共济失调毛细血管扩张和Rad3相关激酶(ataxia telangiectasia and Rad3-related,ATR)是DDR核心的关键激酶,负责感知复制应激(replica-tion stress,RS)并将其信号传导至S和G2/M检查点以启动DNA修复。在肿瘤细胞中G1检查点缺失和癌基因的激活,导致癌症细胞更多进入RS增加的S期。因此,肿瘤细胞更加依赖S和G2/M检查点,使其成为一个有吸引力的靶点。ATR抑制剂是目前抗肿瘤药物开发的热点,部分ATR抑制剂目前已经进入临床试验阶段。本综述旨在总结支持ATR抑制剂作为单药以及与化疗、放疗和新型靶向药物(如PARP抑制剂)联合使用的临床试验数据,并讨论目前ATR抑制剂开发和生物标志物探索中面临的挑战。
文摘Objective:Anlotinib hydrochloride is a multitarget tyrosine kinase inhibitor that targets vascular endothelial growth factor receptor,fibroblast growth factor receptor,platelet-derived growth factor receptor,c-Kit,and c-MET;therefore,it exhibits both antitumor and anti-angiogenetic activities.A phase III trial has shown that anlotinib improved progression-free survival(PFS)and overall survival(OS)in patients with advanced non-small cell lung cancer(NSCLC),who presented with progressive disease or intolerance after standard chemotherapy.This study aimed to analyze the characteristics of patients receiving anlotinib treatment to determine the dominant populations who are fit for the treatment.Methods:Data were collected from March 2015 to January 2017 from a randomized,double-blind,placebo-controlled,multicenter,phase III trial of anlotinib(ALTER0303).A total of 437 patients were enrolled and randomly allocated(2:1)to the anlotinib and placebo groups.Kaplan–Meier analysis and log-rank test were performed to compare PFS and OS.Cox proportional hazards model was adopted for multivariate prognostic analysis.Results:Multivariate analysis indicated that high post-therapeutic peripheral blood granulocyte/lymphocyte ratio and elevated alkaline phosphatase levels were independent risk factors for PFS.Meanwhile,elevated thyroid-stimulating hormone,blood glucose,and triglyceride levels;hypertension;and hand–foot syndrome were independent protective factors of PFS.High posttherapeutic peripheral blood granulocyte/lymphocyte ratio,an Eastern Cooperative Oncology Group(ECOG)score≥2,and the sum of the maximal target lesion length at baseline were independent risk factors of OS,and hypertriglyceridemia was an independent protective factor of OS.Conclusions:This study preliminarily explored the possible factors that affected PFS and OS after anlotinib treatment in patients with advanced refractory NSCLC,and the baseline characteristics of the therapeutically dominant populations were then identified.
基金This work was supported by grants from the National Natural Science Foundation of China(Grant No.81802296)Natural Science Foundation of Tianjin(Grant No.18JCQNJC82500)Tianjin Municipality Science and Technology Commission Projects(Grant No.12ZCDZSY15600).
文摘Objective:Lymphatic metastasis is one of the leading causes of malignancy dispersion in various types of cancer.However,few anti-lymphangiogenic drugs have been approved for clinical use to date.Therefore,new therapies to block lymphangiogenesis are urgently required.Methods:Immunohistochemistry,immunofluorescence,Western blot,migration assays,and lymphangiogenesis and lymphatic metastasis assays were used.Results:Anlotinib,a receptor tyrosine kinase inhibitor,suppressed the rate of new metastatic lesions(31.82%in the placebo arm and 18.18%in the anlotinib arm)in patients with advanced lung adenocarcinoma who were enrolled in our ALTER-0303 study.D2-40+-lymphatic vessel density was strongly correlated with disease stage,metastasis,and poor prognosis in 144 Chinese patients with lung adenocarcinoma.In mice bearing A549EGFP tumors,tumor lymphatic vessel density,tumor cell migration to lymph nodes,and the number of distant metastatic lesions were lower in the anlotinib group than in the controls.Anlotinib inhibited the growth and migration of human lymphatic endothelial cells(hLECs)and lymphangiogenesisin vitro andin vivo.Treatment of hLECs with anlotinib downregulated phosphorylated vascular endothelial growth factor receptor 3(VEGFR-3).Conclusions:Anlotinib inhibits lymphangiogenesis and lymphatic metastasis,probably through inactivating VEGFR-3 phosphorylation.The results indicate that anlotinib may be beneficial for treatment in avoiding lymphangiogenesis and distant lymphatic metastasis in lung adenocarcinoma.(Trial registration:ALTER0303;NCT02388919;March 17,2015.)
文摘Objective:Bevacizumab is a recombinant humanized monoclonal antibody that blocks vascular endothelial growth factor(VEGF)with clear clinical benefits.However,overall survival of some cancer types remains low owing to resistance to bevacizumab therapy.While resistance is commonly ascribed to tumor cell invasion induced by hypoxia-inducible factor(HIF),less attention has been paid to the potential involvement of endothelial cells(ECs)in vasculature activated by anti-angiogenic drugs.Methods:Human umbilical vein ECs(HUVECs),bEnd.3 cells,and mouse retinal microvascular ECs(MRMECs)were treated with bevacizumab under conditions of hypoxia and effects on biological behaviors,such as migration and tube formation,examined.Regulatory effects on TGFpi and CD 105(endoglin)were established via determination o f protein and mRNA levels.We further investigated whether the effects of bevacizumab could be reversed using the receptor tyrosine kinase inhibitor anlotinib.Results:Bevacizumab upregulated TGFpi as well as CD 105,a component o f the TGFP receptor complex and an angiogenesis promoter.Elevated CD 105 induced activation of Sm adl/5,the inflammatory pathway and endothelial-mesenchymal transition.The migration ability of HUVECs was enhanced by bevacizumab under hypoxia.Upregulation o f CD 105 was abrogated by anlotinib,which targets multiple receptor tyrosine kinases including VEGFR2/3,FGFR1-4,PD G FRα/β,C-Kit,and RET.Conclusions:Bevacizumab promotes migration and tube formation of HUVECs via activation of the TGFβi pathway and upregulation of CD105 expression.Anlotinib reverses the effects of bevacizumab by inhibiting the above signals.
基金supported by the National Natural Science Foundation of China(No.31770704)Heilongjiang Touyan Innovation Team Program(Tree Genetics and Breeding Innovation Team)the Overseas Expertise Introduction Project for Discipline Innovation(B16010).
文摘Determination of Cas9 cutting efficiency to the target sites is important for genome editing.However,this determination can only be made via an in vitro method,as the purification of Cas protein and synthesis of gRNA are necessary.Here,we developed an in vivo method,called transient CRISPR/Cas editing in plants(TCEP)to determine Cas9 cutting efficiency.The CRISPR/Cas vector for plant transformation mediated by Agrobacterium tumefaciens was constructed as normal.Using the transient transformation method we built,the Cas9 protein and gRNA were transiently expressed and formed a complex to cut its target sites,resulting in dynamic DNA breakage.The broken DNA was quantified using qPCR to measure the efficiencies of Cas9 cutting.We studied the Cas9 cutting efficiencies to different target sites in Betula platyphylla and Populus davidiana×P.bolleana plants using TCEP and an in vitro method.The results of TCEP were consistent with those of the in vitro method,suggesting that the TCEP method is reliable in determining cutting efficiency.Additionally,using the TCEP method,we showed that both heat and sonication treatment significantly improved CRISPR/Cas efficiency.Therefore,the TCEP method has broad application value and can not only be used to analyze the CRISPR/Cas efficiency but also to determine the factors involved in Cas9 cutting.
